Mal peg scm

Manufactured by Creative PEGWorks

Sourced in New Caledonia, United States

Mal-PEG-SCM is a heterobifunctional PEG reagent that contains a maleimide group and a succinimidyl carbonate group. It can be used for the covalent attachment of PEG to proteins or other biomolecules that contain free thiol or amine groups.

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Lab products found in correlation

Chelex 100 resin, by Merck Group (2 mentions) Lobind tube, by Eppendorf (1 mentions) Slide a lyzer g2 dialysis cassette, by Thermo Fisher Scientific (1 mentions) Afm tips, by Bruker (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) 1 dodecanethiol, by Merck Group (1 mentions) Pd 10 size exclusion column, by GE Healthcare (1 mentions) Fshr mab, by R&D Systems (1 mentions) D luciferin, by GoldBio (1 mentions) P scn bn nota, by Macrocyclics (1 mentions) Traut s reagent, by Thermo Fisher Scientific (1 mentions) Cck 8 kit, by Beyotime (1 mentions) Trypsin, by Beyotime (1 mentions)

6 protocols using mal peg scm

Synthesis of Bifunctional Peptide Conjugate

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All the chemical reactions were performed in low protein binding tubes (LoBind tubes, Eppendorf, Hamburg, Germany). Bicyclic PDGFβR-recognizing peptide (CSRNLIDC-GGGDGG-CSRNLIDC with C”C disulfide bonds, BiPPB, 2223 Da) [18 (link)] and peptidomimetic IFNγ (mimIFNγ, 4689 Da) modified with S-acetyl thioacetate were custom-prepared by Genosphere (Paris, France). To synthesize the construct, BiPPB (0.112 μmol) was reacted with 0.337 μmol of Maleimide-PEG-succinimidyl carboxy methyl ester (Mal-PEG-SCM, 2 KDa, Creative PEGworks, Winston-Salem, NC) for 3 h. Thereafter, excess of Mal-PEG-SCM was blocked with lysine (0.337 μmol) for 1 h. Subsequently, the prepared BiPPB-PEG-Mal was reacted overnight with mimIFNγ-ATA in the presence of deacetylating reagent (0.1M hydroxylamine, 25mM EDTA in PBS pH 7.2). The prepared mimIFNγ-PEG-BiPPB (mimγ-BiPPB [Fibroferon]) conjugate (8.9 KDa) was extensively dialyzed against PBS using 7 kDa dialysis membrane (Thermo Scientific, Rockford, IL).

Poosti F., Bansal R., Yazdani S., Prakash J., Beljaars L., van den Born J., de Borst M.H., van Goor H., Hillebrands J.L, & Poelstra K. (2016). Interferon gamma peptidomimetic targeted to interstitial myofibroblasts attenuates renal fibrosis after unilateral ureteral obstruction in mice. Oncotarget, 7(34), 54240-54252.

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K+ Nanosensor Surface Modification and Peptide Conjugation

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To modify the surface of the K⁺ NS, Bifunctional Polyethylene Glycol (MAL-PEG-SCM, 2 kDa, Creative PEGWorks; 8 mg) was added into K⁺ NS in PBS (pH 7.4; 50 mg/2.5 mL). After 30 min of stirring, it was washed with PBS using an Amicon Ultra-15 Centrifugal Filter (100 kDa) and an F3 Peptide (KDEPQRRSARLSAKPAPPKPEPKP-KKAPAKKC, RS Synthesis; 22 mg) was added and stirred overnight. Cysteine (1.26 mg) was added and stirred for 2 h to deactivate unreacted maleimide groups. The nanosensor solution was washed with water (5×) then concentrated before usage. To confirm F3 conjugation onto the K⁺ NS, a separate batch was synthesized where the F3-peptide was preconjugated with NIR-797. The absorbance of NIR-797 was still visible after the purification.

Lee C.H., Folz J., Zhang W., Jo J., Tan J.W., Wang X, & Kopelman R. (2017). Ion-Selective Nanosensor for Photoacoustic and Fluorescence Imaging of Potassium. Analytical chemistry, 89(15), 7943-7949.

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Synthesis of Mimetic IFNγ Conjugates

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Example 5

Mimetic IFNγ-PEG-BiPPB Conjugate:

0.111 μmol Bicyclic PDGFR recognizing peptide (BiPPB, 2223.2 Da, Genosphere Biotechnologies) was coupled with 0.337 μmol maleimide-PEG-succinimidyl carboxy methyl ester (Mal-PEG-SCM, 2 KDa, Creative PEGworks) for 3 hrs. Thereafter, lysine (0.337 μmol) was added to block free groups of Mal-PEG-SCM. After 1 hr of reaction, the synthesized product BiPPB-PEG-MAL (0.112 μmol) was reacted with 0.56 μmol of Mimetic IFNγ-ATA (4689 Da, Genosphere Biotechnologies) in the presence of deacetylating reagent for overnight at room temperature. Finally, the synthesized Mimetic IFNγ-PEG-BiPPB conjugate (8828.2 Da) was extensively dialyzed against PBS using 7 KDa slide-a-lyzer G2 dialysis cassettes (Thermo scientific).

Mimetic IFNγ-PEG Conjugate.

0.107 μmol Mimetic IFNγ-ATA (4689 Da) was reacted with 0.321 μmol of Poly (ethylene glycol)-succinimidyl α-methylbutanoate (mPEG-SMB, 2 KDa, Nektar therapeutics) for 2 hrs and subsequently the product was dialyzed extensively.

a) Mimetic IFNγ-PEG-BiPPB

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b) Mimetic IFNγ-PEG

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US10442845B2. Interferon analogs (2019-10-15). BiOrion Technologies B.V. [NL]. Inventors: Klaas Poelstra [NL], Jai Prakash [NL], Eleonora Beljaars [NL], Ruchi Bansal [NL].

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Surface Functionalization with Thiol-PEG Linkers

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Tetra-ethylene glycol thiols terminated in hydroxyl (EG4) or amine groups (EG4N) were purchased from Prochimia (Poland). 1-Dodecanethiol (98%) and triethanolamine (TEA) HCl (99%) were purchased from Aldrich (Milwaukee, WI). 11-Aminoundecanethiol was purchased from Dojindo Molecular Technologies. Sulfosuccinimidyl-4-(Nmaleimidomethyl)cyclohexane-1-carboxylate (SSMCC) was purchased from Pierce Biotechnology (Rockford, IL). 10 kDa heterobifunctional PEG linkers (MAL-PEG-SCM) were purchased from Creative PEGWorks. Ethanol (reagent, anhydrous, denatured) used for preparation of thiol solutions and sodium chloride (99.0%) were purchased from Sigma-Aldrich (Milwaukee, WI). Ethanol (anhydrous, 200 proof) used for rinsing was purchased from Decon Laboratories (King of Prussia, PA). Deionized water used in this study had a resistivity of 18.2 MÎ© cm. All chemicals were used as received and without any further purification. The AFM tips used in this study (triangular shaped with radius of curvature of 10 nm) were purchased from Bruker (Camarillo, CA). Silicon wafers were purchased from Silicon Sense (Nashua, NH).

Ma C.D., Acevedo-Vélez C., Wang C., Gellman S.H, & Abbott N.L. (2016). Interaction of the Hydrophobic Tip of an Atomic Force Microscope with Oligopeptides Immobilized Using Short and Long Tethers. Langmuir : the ACS journal of surfaces and colloids, 32(12).

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Synthesis and Characterization of FSHR-Targeted Radiotracers

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AlexaFluor488-labeled rabbit anti-mouse secondary antibodies were purchased from Lampire biological laboratories (Pipersville, PA). p-SCN-Bn-NOTA (i.e., 2-S-(4-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid) was purchased from Macrocyclics, Inc. (Dallas, TX). D-luciferin was acquired from Gold Bio Technology (St Louis, MO). ⁶⁴CuCl₂ was acquired from the University of Wisconsin cyclotron group. SCM-PEG-Mal (i.e. succinimidyl carboxymethyl PEG maleimide, MW: 5 kDa) was purchased from Creative PEG works. FSHR-mAb (catalog number: MAB65591) was purchased from R&D systems (Minneapolis, MN). Chelex 100 resin (50–100 mesh) was acquired from Sigma-Aldrich (St. Louis, MO) and PD-10 size exclusion columns were acquired from GE Healthcare (Piscataway, NJ). Fluorescein isothiocyanate (FITC), Traut’s Reagent, TCEP (i.e. tris(2-carboxyethyl)phosphine), and all the other reagents were purchased from Fisher Thermo Scientific. All the reagents were directly used without further purifications following manufacturer’s instructions. All buffers were prepared from Millipore-grade water and pretreated with the Chelex 100 resin to ensure that the aqueous solution was free of heavy metals.

Yang D., Feng L., Dougherty C.A., Luker K.E., Chen D., Cauble M.A., Banaszak Holl M.M., Luker G.D., Ross B.D., Liu Z, & Hong H. (2016). In Vivo Targeting of Metastatic Breast Cancer via Tumor Vasculature-Specific Nano-Graphene Oxide. Biomaterials, 104, 361-371.

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Targeted Fluorescent Nanoparticle Synthesis

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Manganese (II) acetate (98%), oleic acid, oleylamine, and fluorescein isothiocyanate isomer (FITC) were purchased from Aladdin Inc. (Shanghai, China). DSPE-PEG-NH₂, DSPE-PEG-Mal, and SCM-PEG-Mal and supplied by Creative PEGworks (NC, USA). Core molecule was PEG5000. Traut’s reagent and RGD antibody were bought from Thermo Fisher Inc. (MA, USA). RGD peptide was purchased from Haode Peptide Co., Ltd. (Wuhan, China). Human umbillical vein endothelial cells (HUVECs), human embryonic lung fibroblast cells (IMR-90), human gastric adenocarcinoma cells (SGC-7901), human gastric carcinoma cells (BGC-823), DMEM high glucose medium, and foetal bovine serum (FBS) were purchased from Procell Inc. (Wuhan, China). The CCK-8 kit, DAPI kit, antibiotics and trypsin (0.25%) were ordered from Beyotime Inc. (Shanghai, China). Other chemical reagents were purchased from Sinopharm Corp. (Beijing, China). BALB/c mice and BALB/c-nu/nu mice were purchased from Peking HFK Biotech Corp. (Beijing, China).

Li K., Li P., Wang Y, & Han S. (2020). Manganese-Based Targeted Nanoparticles for Postoperative Gastric Cancer Monitoring via Magnetic Resonance Imaging. Frontiers in Oncology, 10, 601538.

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